Skip to main content
. 2003 Sep;77(17):9632–9638. doi: 10.1128/JVI.77.17.9632-9638.2003

FIG. 3.

FIG. 3.

MVV and CAEV Tat proteins are associated with viral particles. TIGEF were inoculated (MOI, 1) with MVV (A) or CAEV (B and C) or were mock infected. At 24 h postinoculation, the cells were transfected with pTat-Flag-MVV (A) and pTat-Flag-CO (B and C) plasmids. At 48 h posttransfection, the supernatants were collected and virions were harvested from pellets by ultracentrifugation; cell lysates were then obtained following treatment with the lysing solution. (A and B) Proteins from the supernatants (lanes 1 and 3) and cell lysates (lanes 2 and 4) were analyzed by immunoblot assay by using an antibody directed against the FLAG epitope (diluted 10,000 times). Lanes 1 and 2, cells infected with MVV (A) or CAEV (B); lanes 3 and 4, mock-infected cells. (C) Pellets of virions were resuspended and loaded on the top of a linear sucrose gradient. After 16 h of ultracentrifugation, 12 fractions were collected and proteins were extracted and then analyzed by immunoblot assay with an anti-p25 Gag antibody to reveal the major CAEV capsid protein (diluted 1,000 times) or an anti-FLAG antibody (diluted 10,000 times) to reveal the CAEV Tat protein.